Cloning, expression of the psbU gene, and functional studies of the recombinant 12-kDa protein of photosystem II from a red alga Cyanidium caldarium

Hisataka Ohta, Akinori Okumura, Satoshi Okuyama, Ai Akiyama, Masako Iwai, Shizue Yoshihara, Jian Ren Shen, Masaharu Kamo, Isao Enami

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

The encoding extrinsic 12-kDa protein of oxygen-evolving PS II complex from a red alga, Cyanidium caldarium, was cloned and sequenced by means of PCR and a rapid amplification of cDNA ends (RACE) procedure. The gene encodes a putative polypeptide of 154 amino acids with a calculated molecular mass of 16,714 Da. The full sequence of the protein includes two characteristic transit peptides, one for transfer across the chloroplast envelope and another for targeting into the thylakoid lumen. This indicates that the protein is encoded in the nuclear genome. The mature protein consists of 93 amino acids with a calculated molecular mass of 10,513 Da. The cloned gene was successfully expressed in Escherichia coli and the resulting protein was purified, reconstituted to CaCl2-washed PS II complex together with the other extrinsic proteins of 33 and 20 kDa and cyt c-550. The recombinant 12-kDa protein bound completely with the PSII complex, which resulted in a restoration of oxygen evolution equal to the level achieved by binding of the native 12-kDa protein.

Original languageEnglish
Pages (from-to)245-250
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume260
Issue number1
DOIs
Publication statusPublished - Jun 24 1999
Externally publishedYes

Keywords

  • Cyanidium
  • Extrinsic 12-kDa protein
  • Oxygen evolution
  • Photosystem II
  • psbU gene

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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